FIG 1.

(A) Promoter shutdown assay to compare the levels of Nrg1 stability at pH 4 and pH 7 with those of the WT strain containing a copy of MAL2p-NRG1-MYC. A parallel blot was probed with anti-PSTAIRE antibody as a loading control. (B) Reverse transcription-quantitative PCR (qRT-PCR) of NRG1 transcript level after WT cells were grown at pH 4 and pH 7 for 1 h. Quantitative PCR (qPCR) values were normalized to ACT1 values for each sample, and overnight (ON) samples were set to a value of 1. Presented data represent means ± standard errors of the means (SEM) of results from 3 independent experiments. (C) Western blot analysis of Nrg1-myc protein levels after WT cells were inoculated into fresh medium at pH 4 and pH 7 for 1 h. ON, overnight. (D) Chromatin immunoprecipitation (ChIP) analysis of Nrg1 for the promoter of HWP1 after 30 min in YPD medium at pH 4 and pH 7. Presented data represent means ± SEM of results from 3 independent experiments.